Combination of hTERT and bmi-1, E6, or E7 Induces Prolongation of the Life Span of Bone Marrow Stromal Cells from an Elderly Donor without Affecting Their Neurogenic Potential

Abstract

Murine bone marrow stromal cells differentiate not only into mesodermal derivatives, such as osteocytes, chondrocytes, adipocytes, skeletal myocytes, and cardiomyocytes, but also into neuroectodermal cells in vitro. Human bone marrow stromal cells are easy to isolate but difficult to study because of their limited life span. To overcome this problem, we attempted to prolong the life span of bone marrow stromal cells and investigated whether bone marrow stromal cells modified with bmi-1, hTERT, E6, and E7 retained their differentiated capability, or multipotency. In this study, we demonstrated that the life span of bone marrow stromal cells derived from a 91-year-old donor could be extended and that the stromal cells with an extended life span differentiated into neuronal cells in vitro. We examined the neuronally differentiated cells morphologically, physiologically, and biologically and compared the gene profiles of undifferentiated and differentiated cells. The neuronally differentiated cells exhibited characteristics similar to those of midbrain neuronal progenitors. Thus, the results of this study support the possible use of autologous-cell graft systems to treat central nervous system diseases in geriatric patients.

SUPPLEMENTAL MATERIAL

Supplemental material for this article may be found at http://mcb.asm.org/.

ACKNOWLEDGMENTS

We express our sincere thanks to K. Segawa, S. Matsumoto, Y. Okayama, S. Okuyama, and S. Ikeuchi for support throughout the work and to Y. Nakamura, N. Hida, N. Hashimoto, and T. Inomata for providing expert technical assistance. We are grateful to D. A. Galloway (FHCRC, Seattle, Wash.) for pLXSN-16E7 and to Y. Takeuchi (Chester Beatty Laboratories, ICR, United Kingdom) for the FLYA13 cells.

This work was supported in part by a special grant for Advanced Research on Cancer from the Ministry of Education, Culture, Sports, Science, and Technology (MEXT) of Japan to T.K. and A.U.; a grant from MEXT to A.U.; Health and Labour Sciences Research Grants to A.U.; and a grant from the Organization for Pharmaceutical Safety and Research to A.U.